呼吸道菌群失調(diào)加重小鼠過敏性呼吸道炎癥反應(yīng)
發(fā)布時(shí)間：2018-09-13 14:57
【摘要】：【目的】探討呼吸道菌群失調(diào)對(duì)小鼠過敏性呼吸道疾病發(fā)病的影響。【方法】C57BL/6雌性小鼠給予萬古霉素霧化吸入10 d后處死,應(yīng)用16S rRNA高通量測(cè)序技術(shù)分析支氣管肺泡灌洗液(BALF)菌群組成,探索建立小鼠呼吸道菌群失調(diào)模型的方法。建模成功后,應(yīng)用相同的方法建立小鼠呼吸道菌群失調(diào)模型,在此基礎(chǔ)上,通過腹腔注射致敏及霧化吸入卵清蛋白(OVA)激發(fā),誘導(dǎo)小鼠過敏性呼吸道炎癥反應(yīng),計(jì)數(shù)擦鼻頻率,檢測(cè)BALF中細(xì)胞總數(shù)及嗜酸性粒細(xì)胞百分比,肺組織病理分析炎癥反應(yīng)及杯狀細(xì)胞增生情況,ELISA法檢測(cè)血清中IgE水平,BALF中IFN-γ、IL-4、IL-5水平,血清、BALF及腸組織中IL-33水平,從而分析呼吸道菌群失調(diào)對(duì)過敏性呼吸道炎癥反應(yīng)的影響。【結(jié)果】萬古霉素霧化吸入使小鼠呼吸道Bradyrhizobium、Sphingopyxis、Cupriavidus、Pelomonas等菌屬增加,而Akkermansia及Prevotella_6等菌屬顯著降低,發(fā)生明顯的菌群失調(diào)。利用此動(dòng)物模型進(jìn)一步研究發(fā)現(xiàn),呼吸道菌群失調(diào)加重了卵清蛋白誘導(dǎo)的過敏性呼吸道炎癥反應(yīng),表現(xiàn)為擦鼻頻率增加,BALF中細(xì)胞滲出增加、嗜酸性粒細(xì)胞百分比增高,肺組織炎癥及杯狀細(xì)胞增生加重,血清中IgE水平增加。另外,與卵清蛋白組相比,菌群失調(diào)聯(lián)合卵清蛋白組小鼠BALF中Th1型細(xì)胞因子IFN-γ水平降低,Th2型細(xì)胞因子IL-4、IL-5水平升高,發(fā)生更為嚴(yán)重的Th1/Th2平衡失調(diào);BALF中IL-33水平增加,而血清及腸組織中IL-33水平差異無統(tǒng)計(jì)學(xué)意義,表明呼吸道菌群失調(diào)僅影響局部IL-33的產(chǎn)生�！窘Y(jié)論】應(yīng)用萬古霉素霧化吸入成功建立了小鼠呼吸道菌群失調(diào)模型。呼吸道菌群失調(diào)可能通過增加局部IL-33產(chǎn)生,激活Th2及固有樣淋巴細(xì)胞(ILC),誘發(fā)Th1/Th2平衡失調(diào),從而促進(jìn)過敏性呼吸道疾病發(fā)病。
[Abstract]:[objective] to investigate the effect of respiratory tract flora disorder on allergic respiratory diseases in mice. [methods] female C57BL/6 mice were killed by vancomycin aerosol inhalation for 10 days. The composition of (BALF) flora in bronchoalveolar lavage fluid (BALF) was analyzed by 16s rRNA high-throughput sequencing technique. After the successful establishment of the model, the same method was used to establish the model of respiratory tract dysbacteriosis in mice. On this basis, the allergic respiratory inflammation was induced by intraperitoneal injection of ovalbumin (OVA) and atomized inhalation of ovalbumin, and the frequency of nasal rubbing was counted. The total number of cells and the percentage of eosinophils in BALF were detected. The inflammatory reaction and goblet cell proliferation in lung tissue were analyzed. The levels of IgE in serum and IFN- 緯 -IL-4 / IL-5 in BALF and IL-33 in intestinal tissue were detected by Elisa. The results showed that vancomycin atomized inhalation increased the number of Bradyrhizobium,Sphingopyxis,Cupriavidus,Pelomonas in respiratory tract of mice, while Akkermansia and Prevotella_6 decreased significantly, resulting in obvious dysbacteriosis. Using this animal model, we found that the respiratory tract bacterial imbalance aggravated the allergic respiratory inflammation induced by ovalbumin, which showed that the frequency of nasal rubbing increased the exudation of cells and the percentage of eosinophilic granulocytes in BALF, and the percentage of eosinophilic granulocytes in BALF was increased. Pulmonary inflammation and goblet cell proliferation increased, and serum IgE level increased. In addition, compared with ovalbumin group, the level of Th1 type cytokine IFN- 緯 in BALF of mice with dysbacteriosis combined with ovalbumin group decreased, and the level of IL-4,IL-5 of Th2 type cytokines increased, and the IL-33 level in Th1/Th2 balance disorder increased in BALF. However, there was no significant difference in the levels of IL-33 in serum and intestinal tissue, indicating that the respiratory tract flora disorder only affected the production of local IL-33. [conclusion] the model of respiratory tract bacterial flora disorder was successfully established by vancomycin aerosol inhalation in mice. Respiratory dysbacteriosis may promote the pathogenesis of allergic respiratory diseases by increasing the production of local IL-33, activating Th2 and (ILC), of inherent lymphocytes to induce Th1/Th2 imbalance.
【作者單位】： 暨南大學(xué)醫(yī)學(xué)院微生物與免疫學(xué)系;
【基金】：廣東省自然科學(xué)基金(2014A030313370) 暨南大學(xué)科研培育與創(chuàng)新基金(21615420)
【分類號(hào)】：R56
本文編號(hào)：2241503
[Abstract]:[objective] to investigate the effect of respiratory tract flora disorder on allergic respiratory diseases in mice. [methods] female C57BL/6 mice were killed by vancomycin aerosol inhalation for 10 days. The composition of (BALF) flora in bronchoalveolar lavage fluid (BALF) was analyzed by 16s rRNA high-throughput sequencing technique. After the successful establishment of the model, the same method was used to establish the model of respiratory tract dysbacteriosis in mice. On this basis, the allergic respiratory inflammation was induced by intraperitoneal injection of ovalbumin (OVA) and atomized inhalation of ovalbumin, and the frequency of nasal rubbing was counted. The total number of cells and the percentage of eosinophils in BALF were detected. The inflammatory reaction and goblet cell proliferation in lung tissue were analyzed. The levels of IgE in serum and IFN- 緯 -IL-4 / IL-5 in BALF and IL-33 in intestinal tissue were detected by Elisa. The results showed that vancomycin atomized inhalation increased the number of Bradyrhizobium,Sphingopyxis,Cupriavidus,Pelomonas in respiratory tract of mice, while Akkermansia and Prevotella_6 decreased significantly, resulting in obvious dysbacteriosis. Using this animal model, we found that the respiratory tract bacterial imbalance aggravated the allergic respiratory inflammation induced by ovalbumin, which showed that the frequency of nasal rubbing increased the exudation of cells and the percentage of eosinophilic granulocytes in BALF, and the percentage of eosinophilic granulocytes in BALF was increased. Pulmonary inflammation and goblet cell proliferation increased, and serum IgE level increased. In addition, compared with ovalbumin group, the level of Th1 type cytokine IFN- 緯 in BALF of mice with dysbacteriosis combined with ovalbumin group decreased, and the level of IL-4,IL-5 of Th2 type cytokines increased, and the IL-33 level in Th1/Th2 balance disorder increased in BALF. However, there was no significant difference in the levels of IL-33 in serum and intestinal tissue, indicating that the respiratory tract flora disorder only affected the production of local IL-33. [conclusion] the model of respiratory tract bacterial flora disorder was successfully established by vancomycin aerosol inhalation in mice. Respiratory dysbacteriosis may promote the pathogenesis of allergic respiratory diseases by increasing the production of local IL-33, activating Th2 and (ILC), of inherent lymphocytes to induce Th1/Th2 imbalance.
【作者單位】： 暨南大學(xué)醫(yī)學(xué)院微生物與免疫學(xué)系;
【基金】：廣東省自然科學(xué)基金(2014A030313370) 暨南大學(xué)科研培育與創(chuàng)新基金(21615420)
【分類號(hào)】：R56
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